| 重组刚地弓形虫Peroxiredoxin蛋白的纯化及抗血清制备 |
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| 引用本文： | 刘转转,王海龙,孟晓丽,刘红丽,申金雁,殷国荣,原飞. 重组刚地弓形虫Peroxiredoxin蛋白的纯化及抗血清制备[J]. 中国病原生物学杂志, 2011, 0(3) |
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| 作者姓名： | 刘转转 王海龙 孟晓丽 刘红丽 申金雁 殷国荣 原飞 |
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| 作者单位： | 山西医科大学寄生虫学教研室;徐州医学院病原生物与免疫学教研室; |
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| 基金项目： | 国家自然科学基金项目(No.81071374); 徐州医学院科研课题(No.2010KJ16) |
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| 摘 要： | 目的对原核系统表达的重组刚地弓形虫Peroxiredoxin(rTgPrx)蛋白进行镍亲和层析纯化,免疫大鼠,制备抗血清。方法优化rTgPrx表达条件,获得大量的可溶性蛋白,经镍亲和层析法纯化后皮下注射免疫Wistar大鼠,间接ELISA法测定血清抗体效价。结果在表达菌液A600为0.5,IPTG浓度为0.2 mmol/L,37℃诱导10 h时,可溶性rTgPrx表达量较高;用镍亲和层析纯化获得纯的目的蛋白,免疫大鼠后诱导产生高滴度抗体血清,效价达1∶12 800以上。结论镍亲和层析法纯化的rTgPrx具有免疫原性,用该蛋白免疫大鼠可获得高效价的抗rTgPrx血清。
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| 关 键 词： | 刚地弓形虫 Peroxiredoxin 蛋白纯化 抗血清 |
Purification of recombinant peroxiredoxin protein of Toxoplasma gondii and preparation of antisera |
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| LIU Zhuan-zhuan,WANG Hai-long,MENG Xiao-li,LIU Hong-li,SHEN Jin-yan,YIN Guo-rong,YUAN Fei. Purification of recombinant peroxiredoxin protein of Toxoplasma gondii and preparation of antisera[J]. Journal of Pathogen Biology, 2011, 0(3) |
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| Authors: | LIU Zhuan-zhuan WANG Hai-long MENG Xiao-li LIU Hong-li SHEN Jin-yan YIN Guo-rong YUAN Fei |
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| Affiliation: | LIU Zhuan-zhuan1,2,WANG Hai-long1,MENG Xiao-li1,LIU Hong-li1,SHEN Jin-yan1,YIN Guo-rong1,YUAN Fei2(1.Department of Parasitology,Shanxi Medical University,Taiyuan 030001,China,2.Department of Microbiology and Immunology,Xuzhou Medical College,Xuzhou 221004,China) |
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| Abstract: | Objective To purify recombinant peroxiredoxin of Toxoplasma gondii expressed in prokaryotic cells with Ni-NTA affinity chromatography and immune rats and prepare antisera.Methods Conditions for expression of rTgPrx were optimized.Large quantities of soluble rTgPrx were purified with Ni-NTA affinity chromatography.Rats were immunized with purified rTgPrx through subcutaneous injection.The antibody was detected by indirect ELISA.Results Soluble rTgPrx was induced by 0.2 mmol/L IPTG administered for 10 h at 37... |
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| Keywords: | Toxoplasma gondii peroxiredoxin protein purification antisera |
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